Human DAPK3 (ZIP Kinase) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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DAPK3 KO cell line available to order. KO validated by. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1.
View Alternative Names
DAP kinase 3, DAP-like kinase, DAPK3_HUMAN, Death associated kinase 3, Death-associated protein kinase 3, Dlk, EC 2.7.11.1, FLJ36473, MYPT1 kinase, ZIP, ZIP kinase isoform, ZIP-kinase, ZIPK, zipper-interacting protein kinase
- WB
Lab
Western blot - Human DAPK3 (ZIP Kinase) knockout HEK-293T cell line (AB266755)
Lanes 1- 2 : Merged signal (red and green). Green - ab79422 observed at 53 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab79422 was shown to react with ZIP Kinase in wild-type HEK-293T cells in western blot. The band observed in CRISPR/Cas9 edited cell line ab266755 (CRISPR/Cas9 edited cell lysate ab257407) lane below 53kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HEK-293T and DAPK3 CRISPR/Cas9 edited HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab79422 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at a 1 in 5000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-ZIP Kinase antibody [EPR1635] (<a href='/en-us/products/primary-antibodies/zip-kinase-antibody-epr1635-ab79422'>ab79422</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
DAPK3 CRISPR/Cas9 edited HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human DAPK3 (ZIP Kinase) knockout HEK-293T cell line (ab266755)
Predicted band size: 53 kDa
Observed band size: 53 kDa
false
- WB
Lab
Western blot - Human DAPK3 (ZIP Kinase) knockout HEK-293T cell line (AB266755)
Lanes 1- 2 : Merged signal (red and green). Green - ab51602 observed at 53 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab51602 was shown to react with ZIP Kinase in wild-type HEK-293T cells in western blot. The band observed in CRISPR/Cas9 edited cell line ab266755 (CRISPR/Cas9 edited cell lysate ab257407) lane below 53kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HEK-293T and DAPK3 CRISPR/Cas9 edited HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab51602 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-ZIP Kinase antibody [EPR1636Y] (<a href='/en-us/products/primary-antibodies/zip-kinase-antibody-epr1636y-ab51602'>ab51602</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
DAPK3 CRISPR/Cas9 edited HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human DAPK3 (ZIP Kinase) knockout HEK-293T cell line (ab266755)
Predicted band size: 53 kDa
Observed band size: 53 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human DAPK3 (ZIP Kinase) knockout HEK-293T cell line (AB266755)
Homozygous : 1 bp insertion in exon 1
Reactivity data
Product details
Recommended control: Human wild-type HEK293T cell line (ab255449). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
We will provide viable cells that proliferate on revival.
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Handling procedures
Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
ZIP Kinase regulates apoptosis and autophagy impacting cell survival and homeostasis. It does not act in isolation but is part of a complex with other kinases and regulatory proteins. ZIP Kinase interacts with cytoskeletal proteins to mediate cell contraction and motility indicating its involvement in both normal cellular dynamics and stress responses.
Pathways
ZIP Kinase is involved in the apoptosis and autophagy pathways. Within the apoptosis pathway ZIP Kinase partners with proteins like DAPK1 influencing stress and injury responses. In the autophagy pathway it helps in regulating cellular degradation processes. These pathways allow ZIP Kinase to integrate signals that manage cell fate decisions collaboratively modulating cellular life and death.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
Primary Antibodies
AB210528
Anti-ZIP Kinase antibody [EPR18809-86]
RabMAb|Recombinant|KO Validated|Trial Size
![Western blot - Anti-ZIP Kinase antibody [EPR18809-86] (AB210528)](https://content.abcam.com/products/images/zip-kinase-antibody-epr18809-86-ab210528--western-blot-img17898.jpg)
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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